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Extraction of Uracil and Dihydrouracil from Human Serum Using Strata-X PRO SPE Using Kinetex 2.6 μm PS C18 (ESI +Ve mode)

Sample Preparation Details

Part Number :8E-S536-TGA
Description :Strata-X PRO
Condition 1 :1 mL of ACN
Sample Pretreatment :Sample Pre-treatment: • Add 100 µL Serum to tube • Add 900 µL of chilled (~0°C) ACN to the serum while vortexing • Centrifuge samples at 6000 rpm for 5 minutes and transfer supernatant
Load :supernatant (collected from pre-treatment)
Evaporate: :Evaporate to dryness
Reconstitute :100 µL of initial mobile phase
Condition 1 mL of ACN
Load supernatant (collected from pre-treatment)
Vacuum Apply and collect extract
Dry Down Sample under N2 and heat at 45 °C
Reconstitute 100 µL of initial mobile phase

Application Detail (App ID: 25777)

Part No:00F-4780-Y0
Dimensions:150 x 3 mm ID
Elution Type:Gradient
Elution A:0.1%Formic acid in Water
Elution B: Methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 93 7
2 12 93 7

Flow Rate:0.2 mL/min
Col. Temp: 25 °C
Detection: LC/MS/MS @ 0.0000000000 m/z (mass per charge) (25°C)
Note:Add 100 µL Serum (doubly stripped) to tube. Add 900 µL of chilled (~00C) ACN to the serum while vortexing. Centrifuge samples at 6000 rpm for 5 minutes and transfer supernatant
Analyte diluted with initial mobile phase

Analyte Details

Analyte Detail not available

1
5,6 Dihydrouracil (UH2)
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2
Uracil
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3
Uracil 1,3-15N2 (+Ve IS)
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